Primers used for amplification of direct PCR for algC and RT-PCR of internal regions of algC are listed in Table 1.
To confirm the transcription of algC, two internal regions of algC cDNA (1360 bp and 463 bp) were amplified using upstream primer 363F and CD1701R and internal nested primers 1260 nest1F and CD1701R, respectively (Table 1).
To compare gene expression pattern of algC in biofilm versus planktonic A.
Relative Quantification of algC Expression Pattern by Real-Time PCR.
Statistical Correlation of Biofilm Formation with algC Gene Expression.
Amplified algC was purified using gel purification kit (Bangalore GeNei, India) and ligated into pGEMT-Easy vector (Promega, USA) to produce resultant recombinant plasmid pGEalgCA7.
For quantitative biofilm augmentation in the algC clones, biofilms were formed in microtitre plates and quantified as per methods described earlier .
baumannii AIIMS 7 algC gene sequence is JF701279 and AEC46864 for the protein (PMM/PGM) sequence.
Elevated Levels of algC Gene Transcription during Biofilm Mode of Growth.
Biofilm Growth Pattern Correlates with Quantitative algC Gene Expression.
Name Nearest city/ Ab- Type of town bre- collection via- tion American Legion Golf Course Wausau ALGC
Fairway Big Stone Country Club Three Lakes BSCC Fairway Blackhawk Country Club Madison BCC Green Durand Golf Course Durand DGC Fairway Eau Claire Country Club Eau Claire ECCC Green Elks Golf Course Ashland EGC Green Hillcrest Golf & Country Club Altoona HGCC Fairway & Green Lake Forest Golf Course Eagle River LFGC Fairway Luck Golf Course Luck LGC Fairway Maple Bluff Country Club Madison MBCC Green Marshfield Country Club Marshfield MCC Fairway & Green Mellen Golf Course Mellen MeGC Fairway Ojibwa Golf Course Chippewa Falls OGC Fairway Rib Mountain Golf Course Wausau RMGC Fairway SeutryWorld Golf Course Stevens Point SWGC Green St.